The bioterrorism attacks of 2001 highlighted the acute need for rapid, ultra-sensitive, cost-effective, and user-friendly tests to simultaneously screen for multiple types of biowarfare agents at the point of exposure. The goal of the proposed project is to demonstrate the potential of the MultiPath TM test to meet this important need. The MultiPath TM test is designed to rapidly detect multiple disparate agents with the ultra-sensitivity of a nucleic acid amplification test but with the cost-effectiveness and user-friendliness of over-the-counter immunoassay "strip" tests. The test's novel capabilities arise from melding a unique combination of proprietary and existing technologies. The MultiPath TM test uses high-intensity target-specific HiLiter TM microbead labels which produce a signal so intense that individual microscopic beads are easily detected by visual inspection of exposed instant film. The labeling and detection strategy enhances cost-effectiveness and portability by eliminating the need for a light source, imaging optics, photodetectors, and the supporting electrical power sources. Preliminary work described in this proposal indicates the feasibility of key elements of this approach. The proposed project will demonstrate the feasibility of the MultiPath TM test using three model targets (a bacterium, virus, and protein) that emulate the spectrum of targets that might be used in biowarfare. The Specific Aims of the proposal are to (1) demonstrate the use of high-intensity HiLiter TM microbeads for the three model targets; (2) demonstrate the efficacy of a fluidics system that is novel and powerful, yet simple and inexpensive; (3) demonstrate the feasibility of the MultiPath TM test imaging system comprising an instant film detection unit, a scanner, and imaging software; and (4) demonstrate the feasibility of integrating this system to scan samples containing low levels of targets. The Phase 1 results, besides demonstrating the feasibility of the MultiPath TM test, will support the Phase 2 goals which are to develop a MultiPath TM test prototype and show its efficacy in comparisons to "gold standard" tests in the context of "real world" samples. Competitive advantages of the MultiPath TM test include the intellectual property covering key features of the test, an unprecedented feature list of high-end results and customer-friendly attributes, and straightforward commercialization path. Besides addressing the large and growing bioterrorism market, the MultiPath TM test addresses significant unmet testing needs in healthcare (infectious disease, cancer, cardiovascular disease) and in industrial microbiology (food, beverage, pharmaceutical manufacturing). The value that the MultiPath TM test will bring to both the primary and secondary markets is expected to attract large corporations as partners to support commercial development, marketing, and distribution. [unreadable] [unreadable]